Intermediates for 2,3-diamino-1,5-iminosugars

ABSTRACT

Novel derivatives of 1-deoxynojirimycin are disclosed which have amino or azido substituents at C-2 and/or C-3. These compounds are useful inhibitors of lentiviruses. Methods of chemical synthesis of these derivatives and intermediates therefor are also disclosed.

This is a DIVISION, of application Ser. No. 08/169,233, filed Dec. 20,1993, now U.S. Pat. No. 5,391,746, which is a division of Ser. No.08/002,380, filed Jan. 8, 1993, now U.S. Pat. No. 5,334,717, which is aDivision of application Ser. No. 07/861,696, filed Apr. 1, 1992, nowU.S. Pat. No. 5,206,251.

BACKGROUND OF THE INVENTION

This invention relates to novel derivatives of1,5-dideoxy-1,5-imino-D-glucitol having amino or azido substituents atC-2 and/or C-3, and, more particularly, to the chemical synthesis ofthese derivatives and intermediates therefor, and to their method ofinhibiting viruses such as lentiviruses.

1,5-dideoxy-1,5-imino-D-glucitol (deoxynojirimycin or DNJ) and itsN-alkyl and O-acylated derivatives are known inhibitors of viruses suchas human immunodeficiency virus (HIV). See, e.g., U.S. Pat. Nos.4,849,430; 5,003,072; 5,030,638 and PCT Int'l. Appln. WO 87/03903.Several of these derivatives also are effective against other virusessuch as HSV and CMV as disclosed in U.S. Pat. No. 4,957,926. In somecases antiviral activity is enhanced by combination of the DNJderivative with other antiviral agents such as AZT as described in U.S.Pat. No. 5,011,829. Various of these DNJ derivative compounds areantihyperglycemic agents based on their activity as glycosidaseinhibitors. See, e.g., U.S. Pat. No. 4,182,763, 4,533,668 and 4,639,436.The 2-acetamide derivatives of DNJ also are reported to be potentglycosidase inhibitors by Fleet et al., Chem. Lett. 7, 1051-1054 (1986);and Kiso et al. J. Carbohydr. Chem. 10, 25-45 (1991).

Notwithstanding the foregoing, the search continues for the discoveryand novel synthesis of new and improved antiviral compounds.

BRIEF DESCRIPTION OF THE INVENTION

In accordance with the present invention, novel derivatives of1,5-dideoxy-1,5-imino-D-glucitol having amino or azido substituents atC-2 and/or C-3 are provided. According to another embodiment of theinvention, novel methods of chemical synthesis of these DNJ derivativesand their intermediates are provided. The novel DNJ derivatives andvarious of their intermediates have useful antiviral activity asdemonstrated against lentivirus.

The novel C-2 and/or C-3 amino or azido substituted derivatives of1,5-dideoxy-1,5-imino-D-glucitol can be represented by the followinggeneral structural Formula I: ##STR1## wherein R=H, alkyl, and aralkyl;

X₁ =OH, N₃, NH₂, NHR₁, NR₂ and NHCOR₃ ;

X₂ =OH, N₃ and NH₂, provided that when X₂ is N₃ or NH₂, X₁ is OH or NH₂,and provided further that at least one of X₁ and X₂ is not OH;

R₁,R₂ =alkyl; and

R₃ =H, alkyl.

In Formula I, the alkyl moieties in the R, R₁, R₂ and R₃ substituentspreferably are straight chain or branched alkyl groups or cycloalkylgroups which preferably have from one to about 8 carbon atoms in R₁, R₂and R₂, e.g., methyl, ethyl, propyl, isopropyl, n-butyl, iso-butyl,sec.-butyl. tert-butyl, pentyl, hexyl, heptyl, octyl, 2-ethylbutyl,2-methylpentyl, cyclopentyl and cyclohexyl, and from one to about 18carbon atoms in R, e.g., dodecyl, octadecyl or any of the above groups.

Also in Formula I, the aryl moieties in the R substituents preferablyare phenyl and substituted phenyl, e.g. benzyl, 4,fluorophenyl or3-methoxyphenyl.

Preferred compounds of Formula I are the following:

2-Azido Derivatives of DNJ

2-Azido-1,2,5-trideoxy-1,5-imino-D-glucitol

2-Azido-1,5- (butylimino) -1,2,5-trideoxy-D-glucitol

2-Azido-1,5-[(2-ethylbutyl)imino]-1,2,5-trideoxy-D-glucitol

2-Azido-1,5-[(4,4,4-trifluorobutyl)imino]-1,3,5-trideoxy-D-glucitol

2-Amino Derivatives of DNJ

2-Amino-1,2,5-trideoxy-1,5-imino-D-glucitol

2-Amino-1,5-(butylimino)-1,2,5-trideoxy-D-glucitol

2-Amino-1,5-[(2-ethylbutyl)imino]-1,2,5-trideoxy-D-glucitol

2-Amino-1,5-[(4,4,4-trifluorobutyl)imino]-1,2,5-trideoxy-D-glucitol

1,5-(Butylimino)-1,2,5-trideoxy-2-(dimethylimino)-D-glucitol

1,5-(Butylimino)-1,2,5-trideoxy-2-(methylamino)-D-glucitol

1,5(Butylimino)-1,2,5-trideoxy-2-[(1-oxobutyl)amino]-D-glucitol

1,5(Butylimino)-1,2,5-trideoxy-2-[(1-oxobutyl)amino]-D-glucitol,tributanoate

3-Amino Derivatives of DNJ

3-Amino-1,3,5-trideoxy-1,5-imino-D-glucitol

2,3-Diamino-1,5-(butylimino) -1,2,3,5-tetradeoxy-D-glucitol

The novel synthesis of compounds of Formula I comprises the formation ofstructural modifications at C2 and C3 of DNJ and the nucleophilicopening of N-carboalkoxy-2,3-anhydro-DNJ.

In accordance with a preferred embodiment of the invention, thecompounds of Formula I can be chemically synthesized by the sequence ofreactions shown in the following generic Reaction Schemes A, D and F inwhich the Roman numerals in parentheses refer to the compounds definedby the generic formula shown above said numbers. R₁ can be any alkyl oraryl group such as illustrated by the reactants and products describedhereinafter. ##STR2##

The foregoing Reaction Scheme A comprises the following general reactionsteps:

(a) The starting material, DNJ (I), is N-acylated with an acylatingagent to form a carbamate derivative of DNJ (II);

(b) The hydroxyls at C-4 and C-6 are protected with a hydroxylprotecting agent by acetalization or ketalization to form an acetal orketal (III);

(c) The hydroxyl at C-2 is protected by regioselective sulfonylationwith a sulfonylating agent at C-2 to give the 2-sulfonated intermediate(IV);

(d) A 2,3-anhydro derivative is formed by epoxidation at C-2 and C-3 togive the epoxide intermediate (V);

Epoxide intermediate (V) is used for synthesis of 2-azido and 2-aminoderivatives of DNJ in the following steps of Reaction Scheme A orretained for synthesis of 3-amino derivatives of 1,5-imino-D-altritol inReaction Scheme H.

(e) The epoxide intermediate (V) is opened by nucleophilic attack at C-2and C-3 such as with an azide to give a mixture of azido derivatives(VI) and (VII);

Azido derivative (VII) is retained for synthesis of 3-azido and 3-aminoderivatives of DNJ in Reaction Scheme D.

Azido derivative (VI) is used for synthesis of 2-azido and 2-aminoderivatives of DNJ in the following steps of Reaction Scheme A orretained for synthesis of 2,3-diamino derivatives of DNJ in ReactionScheme F.

(f) The N-carbamate group in azido derivative (VI) is removed to giveintermediate (VIII).

(g) Intermediate (VIII) is N-alkylated to give the divergentintermediate (IX) which can be used to prepare the final 2-azido or2-amino derivatives of DNJ.

(h) The hydroxyl protecting group at C-4 and C-6 of intermediate (IX) isremoved by cleavage of acetal or ketal to give the desired novelantiviral 2-azido derivatives of DNJ (X),

(i) The 2-azido group in intermediate (IX) is reduced to the 2-aminogroup to give intermediate (XI);

(j) The hydroxyl protecting group at C-4 and C-6 of intermediate (XI) isremoved by cleavage of acetal or ketal to give the desired novelantiviral 2-amino derivatives of DNJ (XII).

N-Acylation of DNJ (I) in step (a) can be carried out by conventionalN-acylation procedures well known to those skilled in the art. Suitablegeneral procedures for acylation of amines are described in U.S. Pat.No. 5,003,072; March, J. in Advanced Organic Chemistry, Wiley, New York,1985; Patai, S. (Ed.) in The Chemistry of Amides, Wiley, New York, 1970.For example, DNJ is N-acylated to form carbamate or thiocarbamate usinga variety of reagents such as chloroformates (e.g., methylchloroformate, ethyl chloroformate, vinyl chloroformate, benzylchloroformate) or dicarbonates (e.g., di-tert-butyl dicarbonate). Thereaction of DNJ (I) with anhydrides, chloroformates or dicarbonates ispreferentially carried out by dissolving in one or more of polar, proticsolvents (such as water, methanol, ethanol) and in the presence of abase (e.g, potassium carbonate, lithium carbonate, sodium carbonate,cesium carbonate, triethylamine, pyridine, 4-dimethylaminopyridine,diisopropylethylamine, 1,8-diazabicyclo[5,4,0]undec-7-ene). N-Acylationis preferentially carried out by reacting DNJ (I) with alkyl or arylchloroformate in solvents such as DMF or aqueous sodium bicarbonate at20°-50° C. to give the product (II).

Protection of the hydroxyl groups at C-4 and C-6 in step (b) to giveacetal or ketal derivative (III) can be carried out by conventionalhydroxyl protection procedures such as those described, e.g., in U.S.Pat. No. 5,003,072 and in Green, T. W., Protective Groups in OrganicSynthesis, Wiley, New York, 1981. The cyclic acetals and ketals areformed by the reaction of 4,6-dihydroxy compound (II) with an aldehydeor a ketone in the presence of an acid catalyst. Illustrative carbonyl(or carbonyl equivalents such as dimethyl acetal or dimethyl ketal)compounds useful in this reaction are acetone, acetaldehyde, methylphenyl ketone, benzaldehyde, 4-methoxybenzaldehyde,2,4-dimethoxybenzaldehyde, 4-dimethylaminobenzaldehyde,2-nitrobenzaldehyde, 2,2,2-trichloroacetaldehyde (chloral) andacetophenone. The acid catalysts suitable for this reaction are, e.g.,para-toluene sulfonic acid, cat. HCl, cat. sulfuric acid, FeCl₃, ZnCl₂,SnCl₂ and BF₃ -ether, and the reaction is carried out in the presence ofaprotic solvents such as methylene chloride, 1,2-dimethoxyethane,dioxane, dimethylformamide, dimethylacetamide or dimethylsulfoxide. Thuspara-toluene sulfonic acid is added to a solution of benzaldehydedimethyl acetal in organic medium, e.g., dimethylformamide, and reactedwith N-acyl-DNJ (II) at 20°-65° C. to give the product (III).

The selective protection of the hydroxy group at C-2 in compound (III)in step (c) can be carried out by regioselective tosylation to give thetosylate (IV). For example, compound (III) is conveniently refluxed withdibutyltinoxide in solvents (such as benzene, toluene, xylene, methanolor ethanol and the like) to form a homogeneous solution. The stannyleneintermediate is then reacted with p-toluenesulfonyl chloride to givetosylate (IV). Other sulfonyl chlorides such as benzenesulfonylchloride, 4-bromobenzenesulfonyl chloride, 4-nitrobenzenesulfonylchloride and methanesulfonyl chloride can also be used in this reaction.

The epoxide intermediate (V) is readily prepared in step (d) bytreatment of the sulfonate (IV) with base such as sodium hydride,potassium hydride, lithium hydride, cesium carbonate, potassiumcarbonate and potassium tert-butoxide using solvents such asdimethylformamide, dimethylacetamide, dimethylsulfoxide,dimethoxyethane, tetrahydrofuran, dioxane, diethyl ether, dibutyl etherand tert-butyl methyl ether.

The nucleophilic opening of epoxide intermediate (V) in step (e) ispreferably carried out by heating (50° C.-reflux) a solution of (V) insolvents such as dimethylformamide, dimethylacetamide, 2-methoxyethanol,dimethoxyethane, tetrahydrofuran, dioxane, dibutl ether and tert-butylmethyl ether with sodium azide to give diasteromeric mixture of theproducts (VI) and (VII).

The nitrogen protecting carbamate group in compound (VI) can be easilyremoved in step (f) by base hydrolysis at temperature of 40° to 100° C.to give the intermediate (VIII). Illustrative bases suitable for thisreaction are aqueous sodium hydroxide, lithium hydroxide or potassiumhydroxide with or without the presence of organic solvents such asmethanol, ethanol, ethylene glycol and dioxane. The carbamates can alsobe cleaved by other reagents such as sulfur nucleophiles (e.g., sodiumthiomethoxide and lithium thiopropoxide) or iodotrimethylsilane.

N-Alkylation of intermediate (VIII) can be carried out in step (g) byreductive alkylation procedures using NaCNBH₃, NaBH₄ or alkylaldehyde.Appropriate alkylaldehydes for preparing the corresponding N-alkylderivative compounds (IX) are, e.g., n-propanal, n-butanal, n-pentanal,n-hexanal, n-heptanal and n-octanal. Preferred aldehydes for thisreaction are, e.g., butyraldehyde, 3-phenylpropionaldehyde and2-ethylbutyraldehyde.

Alternatively, N-alkylation can be achieved by reacting intermediate(VIII) with alkylhalide such as benzyl bromide, bromobutane,bromohexane, iodomethane and the like in the presence of a base such astriethylamine, pyridine and diisopropylethylamine. Suitable solvents forthe reaction are, e.g., DMF, dimethylacetamide, dimethylsulfoxide andpyridine. A preferred alkylhalide for the N-alkylation is1-bromo-4,4,4-trifluorobutane.

The acetal or ketal group from the intermediate (IX) can be removed byacid catalyzed hydrolysis in step (h) to give the novel 2-azidoderivatives of DNJ (X). Acids can be used such as trifluoacetic acid(with or without water), aqueous hydrochloric acid, boron trichloride,1N sulfuric acid, 80% acetic acid, with acidic resin (such as Dowex50-W, H⁺) catalytic p-toluenenesulfonic acid in methanol or ethanol at25°-80° C. The benzylidine acetal can also be cleaved usingN-bromosuccinimide and BaCO₃ (or CaCO₃) in carbon tetrachloride or byeletrochemical reduction.

Reduction of the 2-azido group in intermediate (IX) to give the 2-aminointermediate (XI) in step (i) is conveniently carried out byhydrogenation with palladium on carbon. The acetal or ketal group canthen be removed from intermediate (XI) in step (j) by using conditionssimilar to those elaborated in step (h) to give the novel 2-aminoderivatives (XII) of DNJ. When the 4,6-hydroxy protecting group in (XI)is benzylidine acetal, the group may be removed by transferhydrogenation conditions (e.g., heating a solution of (XI) in ethanolwith Pd(OH)₂ and hydrogen donors such as cyclohexene or1,4-cyclohexadiene). The benzylidine group in (XI) can similarly beremoved by using metals (such as Li, Na or K) and liquid ammonia at -70°to -33° C. to give (XII). The benzylidine acetal can also be cleavedusing N-bromosuccinimide and BaCO₃ (or CaCO₃) in carbon tetrachloride orby electrochemical reduction. 2,2,2-Trichloroethylidine acetal can alsobe cleaved by catalytic reduction (H₂, Raney Ni) using aqueous sodiumhydroxide and ethanol.

The following Reaction Schemes B and C show the preferred synthesis of,respectively, the 2-azido and 2-amino derivatives of DNJ (Scheme B) andthe 2-alkylamino and 2-acylamino derivatives of DNJ (Scheme C), in whichthe arabic numerals in parentheses refer to compounds prepared indetailed Examples set forth hereinbelow: ##STR3##

The foregoing Reaction Scheme D, which shows the generic synthesis ofthe 3-azido- and 3-amino-DNJ derivatives of Formula I, comprises thefollowing general reaction Steps, starting with azido derivative (VII)which was prepared in step (e) of Reaction Scheme A:

(a) The free hydroxyl group at C-2 in the starting material, azidoderivative (VII), is oxidized to give ketone (XIII);

(b) The ketone (XIII) is reduced with a reducing agent such as, e.g.,diisobutylaluminum hydride, sodium borohydride and the like, to give themixture of epimeric alcohols (XIV) and (XV);

(c) The N-carbamate in alcohol (XIV) is hydrolytically cleaved to giveintermediate (XVI);

(d) Intermediate (XVI) is N-alkylated to give divergent intermediate(XVII) which can be used to prepare the final 3-azido or 3-aminoderivatives of DNJ;

(e) The hydroxyl protecting group at C-4 and C-6 of intermediate (XVII)is removed by cleavage of acetal or ketal to give the desired 3-azidoderivatives of DNJ (XVIII);

(f) The 3-azido group in intermediate (XVII) is reduced to the 3-aminogroup to give intermediate (XIX);

(g) The hydroxyl protecting group at C-4 and C-6 of intermediate (XIX)is removed by cleavage of acetal or ketal to give the desired 3-aminoderivatives of DNJ (XX).

In the foregoing Reaction Scheme D, steps (c) through (g) for thesynthesis of the 3-azido and 3-amino derivatives of DNJ can be carriedout with similar reagents and conditions in a manner analogous to steps(f) through (j) used for the synthesis of the 2-azido and 2-aminoderivatives of DNJ in Reaction Scheme A. In step (a), since the rest ofthe molecule is fully protected, the oxidation of secondary alcohol in(VII) can be successfully carried out by a variety of oxidizing agents.(see, e.g., March, J. in Advanced Organic Chemistry, Wiley, New York,1985; House, H. O. in Modern Synthetic Reactions, Benzamin PublishingCo., Massachusetts, 1972; Augusting, R. L. in Oxidations--Techniques andApplications in Organic Synthesis, Dekker, New York, 1969; W. P.Griffith and S. M. Levy, Aldrichchimica Acta 23, 13 (1990); R. M.Moriarty and O. Prakash J. Org. Chem. 50, 151, 1985; A. Mancuso, D.Swern, Synthesis, 165 (1981); S. Czernecki, C. Georgoulus, C. L. Stevensand K. Vijayakantam, Tetrahedron Lett. 26, 1699 (1985); J. Hersovici, M.J. Egra and K. Antonakis, J. Chem. Soc. Perkin Trans. I, 1967 (1982); E.J. Corey, E. Barrette and P. Margriotis, Tetrahedron Lett. 26, 5855(1985); H. Tomioka, K. Oshima and H. Nozaki, Tetrahedron Lett. 23, 539(1982). Some of the reagents suitable for oxidation of the C-2 hydroxylin compound VII are pyridinium chlorochromate (with or without additivessuch as sodium acetate, celite, alumina, molecular sieves), pyridiniumdichromate, chromium trioxide/pyridine, 2,2'-bipyridiniumchloroacromate, cyclic chromate ester (E. J. Corey, E. Barrette and P.Margriotis, Tetrahedron Lett. 26, 5855 (1985), RuCl₂ (PPh₃)₃-tert-BuOOH, silver carbonate on celite, cerium (IV) ammonium nitrate(with or without sodium bromate), tetra-n-propylammonium perruthenate,activated dimethyl sulfoxide reagents (using DMSO and one of theelectrophilic reagents such as acetic anhydride, trifluoroaceticanhydride, oxalyl chloride, trifluorosulfonic anhydride,dicyclohexylcarbodiimide). Formation of the novel carbonyl compound(XIII) is preferentially carried out by oxidation of the hydroxyl groupat C-2 (VII) with trifluoroacetic anhydride in dimethylsulfoxide (DMSO)using methylene chloride as solvent at -70° to 0° C. followed bytreatment with base such as triethylamine or diisopropylethylamine at-70° to 25° C.

The following Reaction Scheme E shows the preferred synthesis of the3-amino derivatives of DNJ in which the arabic numerals in parenthesesrefer to compounds prepared in detailed Examples set forth hereinbelow.Table 1, below, sets forth the results obtained in proportions ofalcohols (33) and (34) by the reduction of ketone (32) under variousreducing conditions.

                  TABLE 1                                                         ______________________________________                                        Studies on Stereoselective Reduction of 32                                     ##STR4##                                                                      ##STR5##                                                                      ##STR6##                                                                                              Relative                                                                      Yield    Chem. Yield                                 Reducing Agent                                                                            Conditions   (33/34)  (33 + 34, %)                                ______________________________________                                        NaBH.sub.4  THF/MeOH (4/1),                                                                            52/48    67                                                      -15° -0° C.,                                                    30 min                                                            LIBH.sub.3 Me                                                                             -70° to -20° C.,                                                             37/63    20                                          MeLi--LiBr +                                                                              3 hr                                                              BH.sub.3 --Me.sub.2 S                                                         Me.sub.3 Al, t-BuMgCl,                                                                    0-5° C., 5 hr                                                                       85/15    26                                          2,6-dibutyl-4-methyl                                                          phenol                                                                        DIBAL-H     -70° C., 4 hr                                                                       86/14    86                                          (1M soln. in                                                                  toluene)                                                                      ______________________________________                                         ##STR7##

The foregoing Reaction Scheme F, which shows the generic synthesis ofthe 2,3-diamino-DNJ derivatives of Formula I, comprises the followinggeneral reaction steps, starting with azido derivative (VI) which wasprepared in step (e) of Reaction Scheme A:

(a) Azido derivative (VI) is subjected to inversion of configuration atC-3 to give the talo intermediate (XXI);

(b) The free hydroxyl at C-3 of intermediate (XXI) is sulfonated to givesulfonic ester (XXII);

(c) Sulfonic ester (XXII) is subjected to azide displacement withinversion to the gluco configuration at C-3 to give diazido intermediate(XXIII);

(d) The N-carbamate in diazido intermediate (XXIII) is hydrolyticallycleaved to give intermediate (XXIV);

(e) Intermediate (XXIV) is N-alkylated to give divergent intermediate(XXV) which can be used to prepare the final 2,3-diamino or 2,3-diazidoderivatives of DNJ;

(f) The hydroxyl protecting group at C-4 and C-6 of intermediate (XXV)is removed by cleavage of acetal or ketal to give the 2,3-diazidoderivatives of DNJ (XXVI);

(g) The 2,3-diazido groups in intermediate (XXV) are reduced to the2,3-diamino groups to give intermediate (XXVII);

(h) The hydroxyl protecting group at C-4 and C-6 of intermediate (XXVII)is removed by cleavage of acetal or ketal to give the desired2,3-diamino derivatives of DNJ (XXVIII).

In the foregoing Reaction Scheme F, steps (d) through (h) for thesynthesis of the 2,3-diazido and 2,3-diamino derivatives of DNJ can becarried out with similar reagents and conditions in a manner analogousto steps (f) through (j) used for the synthesis of the 2-azido and2-amino derivatives of DNJ in Reaction Scheme A. Reaction steps (a)through (c) involve displacement of hydroxyl at C-3 by an azide groupwith net retention of configuration.

The following Reaction Scheme G shows the preferred synthesis of the2,3-diamino derivatives of DNJ in which the arabic numerals inparenthesis refer to compounds prepared in detailed Examples set forthhereinafter. ##STR8##

The following Reaction Scheme H shows the synthesis of 3-aminoderivatives of 1,5-imino-D-altritol from epoxide intermediate (V) ofReaction Scheme A and, preferably, the epoxide intermediate (5) ofReaction Scheme B. Reaction Scheme H comprises the following reactionsteps in which the arabic numerals in parentheses refer to compoundsprepared in detailed Examples set forth hereinbelow:

(a) The epoxide intermediate (5) is opened by refluxing in alkylaminesuch as N,N-dimethylaminoethylamine or butylamine to give the C-4 andC-6 hydroxyl protected 3-amino derivatives of 1,5-imino-altritol (47)and (48), respectively;

(b) The benzyl carbamate (Z) on the 3-amino derivative (47) is removedby base hydrolysis or by catalytic hydrogenation procedures (H₂ and Pd/Cor H₂ and Pd black) to give intermediate (49); and

(c) The hydroxyl protecting group at C-4 and C-6 of intermediate (49) isremoved by cleavage of acetal or ketal to give the desired novelantiviral 3-amino derivative of 1,5-imino-D-altritol (50). This step canbe carried out in a manner analogous to the acid catalysed hydrolysis toremove the acetal or ketal group from intermediate (IX) in step (h) ofReaction Scheme A. ##STR9##

In standard in vitro tests, the novel compounds of the invention weredemonstrated to have inhibitory activity against visna virus in aconventional plaque reduction assay. Visna virus, a lentivirusgenetically very similar to the AIDS virus, is pathogenic for sheep andgoats. See Sonigo et al., Cell 42, 369-382 (1985); Haase, Nature 322,130-136 (1986). Inhibition of visna virus replication in vitro as auseful model for human immunodeficiency virus (HIV) and its inhibitionby test compounds has been described by Frank et al., AntimicrobialAgents and Chemotherapy 31(9), 1369-1374 (1987).

Inhibition of HIV-1 can be shown by tests involving plating ofsusceptible human host cells which are syncytium-sensitive with andwithout virus in microculture plates, adding various concentrations ofthe test compound, incubating the plates for 9 days (during which timeinfected, non-drug treated control cells are largely or totallydestroyed by the virus), and then determining the number of remainingviable cells using a colorimetric endpoint.

DETAILED DESCRIPTION OF THE INVENTION

The following examples will further illustrate the invention although itwill be understood that the invention is not limited to these specificexamples or the details disclosed therein.

EXAMPLE 1 Preparation of1,5-dideoxy-1,5-[{(phenylmethoxy)carbonyl}imino]-D-glucitol (2)

To a stirred-solution of 1-deoxynojirimycin (100 g, 0.61 mol) insaturated aqueous sodium bicarbonate (1000 ml), benzyl chloroformate(95%, 121 g, 0.67 mol) was added dropwise at room temperature. Afterstirring at room temperature for 18 hr, the solution was extracted oncewith methylene chloride (300 ml) to remove any unreacted benzylchloroformate. The aqueous layer was then extracted several times withethyl acetate to give a total of 2.5-3 liters of the extract. Theorganic layer was then dried (Na₂ SO₄), filtered and concentrated togive (2) a white solid (98.57 g, 54%), mp 101°-2° C. Anal calcd. for C₁₄H₁₉ NO₆ C, 56.56, H, 6.44, N, 4.71 Found C, 56.33, H, 6.38, N, 4.58., ¹H NMR (CD₃ OD) 7.2-7.4 (m, 5H), 5.15 (s, 2H), 4.23 (br m, 1H), 4.05 (brd., J =8 Hz, 1H), 3.87 (dd, J=6, 4 Hz, 1H), 3.78-3.85 (m, 2H), 3.70-3.78(m, 2H), 3.45 (br d, J=8 Hz, 1H).

EXAMPLE 2 Preparation of1,5-dideoxy-1,5-[{(phenylmethoxy)carbonyl}imino]-4,6-O-(R-phenylmethylene)-D-glucitol(3)

A mixture of (2) (98.5 g, 0.33 mol), benzaldehyde dimethyl acetal (65.5g, 0.43 mol) and p-toluenesulfonic acid (1 g) in a round bottom flaskwas dissolved in dimethlformamide (400 ml). The flask was connected to awater aspirator and the reaction was heated to 60°-65° C. for 4 hr. Thereaction mixture was cooled to room temperature and poured into stirredice-water (1200 ml) containing sodium bicarbonate (14 g). The whitesolid formed was filtered, washed with cold water and dried.Recrystallization using hexane/ethyl acetate gave 3 (96.2 g, 54%) aspure white solid, mp 147°-48° C., Anal calcd. for C₂₁ H₂₃ NO₆ C, 65.44,H, 6.02, N, 3.63 Found C, 65.15, H, 5.93, N, 3.49. IR (KBr) 3420, 1715,1450, 1425, 1395, 1380, 1365, 1090 cm⁻¹ ; ¹ H NMR (CD₃ OD) 7.28-7.53 (m,10H), 5.61 (s, 1H), 5.14 (s, 2H), 4.77 (dd, J=11, 4.6 Hz, 1H), 4.38 (t,J=11 Hz, 1H), 4.16 (dd, J=13.4, 4.2 Hz, 1H), 3.5-3.7 (complex m, 3H),3.35 (td, J=11, 4.6 Hz), 2.97 (dd, J=13.4, 9.3 Hz, 1H); ¹³ C NMR (CD₃OD) 156.7, 139.4, 138.0, 129.9, 129.7, 129.3, 129.2, 129.1, 127.6,102.8, 81.9, 77.5, 71.5, 70.6, 68.6, 55.9 and 50.5; MS (CI, NH₃, m/e)386 (M+1).

EXAMPLE 3 Preparation of1,5-dideoxy-1,5-[{(phenylmethoxy)carbonyl}imino]-4,6-O-(R-phenylmethylene)-D-glucitol,2-(4-methylbenzenesulfonate) (4)

A mixture of diol 3 (46.3 g, 0.12 mol) and di-n-butyltin oxide (31.1 g,0.125 mol) in methanol (300 ml) was refluxed for 2 hr. The methanol wasremoved, toluene was added and removed under vaccuum. The residue wasdissolved in methylene chloride (300 ml) and triethylamine (20 ml, 0.144mmol) After cooling to 0° C., p-toluenesulfonyl chloride (25.2 g, 0.132mmol) was added. The reaction was stirred at 0° C. for 30 min and thenwarmed to 20° C. After stirring for 3 hr, the reaction was quenched byadding saturated aqueous sodium bicarbonate. The organic layer wasseparated and washed with water, 0.5M KHSO₄ and water successively. Theorganic layer was dried (Na₂ SO₄), filtered and concentrated. Theresidue was chromatographed (silica gel, hexane/ethyl acetate 7/3) togive pure 4 (50.27 g, 77%) as white solid, mp 115°-17° C., Anal calcd.for C₂₈ H₂₉ NO₈ S: C, 62.32, H, 5.42, N, 2.66 Found C, 62.65, H, 5.40,N, 2.62. ¹ H NMR (CDCl₃) 7.82 (d, J=7.8 Hz, 2H), 7.35-7.50 (m, 10H),7.31 (d, J=7.8 Hz, 2H), 5.51 (s, 1H), 5.12 (s, 2H), 4.76 (dd, J=11.4,4.5 Hz, 1H), 4.38 (ddd, J=9.3, 7.6, 4.8 Hz, 1H), 4.32 (dd, J=11.4, 9.5Hz, 1H), 4.31 (dd, J=13.6, 4.8 Hz, 1H), 3.78 (dt, J=2.6, 9.4Hz, 1H),3.59 (t, J=9.4 Hz, 1H), 3.26 (ddd, J=11.4, 9.4, 4.5 Hz, 1H), 3.04 (dd,J=13.6, 9.3 Hz, 1H) 2.63 (d, J=2.6 Hz, 1H), 2.41 (s, 3H); ¹³ C NMR(CDCl₃) 154.8, 145.2, 137.0, 135.8, 133.2, 129.8, 129.3, 128.7, 128.4,128.3, 128.1, 126.2, 101.8, 79.9, 78.1, 73.9, 69.2, 67.8, 54.2, 47.1 and21.7; MS (m/e) 546 (M+Li).

EXAMPLE 4 Preparation of2,3-anhydro-1,5-dideoxy-1,5-[{(phenylmethoxy)carbonyl}imino]-4,6-O-(R-phenylmethylene)-D-mannitol(5)

Sodium hydride (2.79 g, 60% dispersion in mineral oil, 69.66 mol) wasplaced in a flask under argon and washed three times with dry hexane.The residue was suspended in dry THF (300 ml) and to this a solution of4 (37.6 g, 69.66 mmol) in THF (100 ml) was added slowly. After stirringfor 18 hr, the reaction was quenched by adding water. The organic layerwas extacted with ethyl acetate and washed with saturated aqueous sodiumbicarbonate and brine. After drying (sodium sulfate) and filteration,the organic layer was concentrated and recrystallized using cyclohexaneto give pure 5 (19.2 g, 75%) as white solid, mp 104°-5° C. Anal calcdfor C₂₁ H₂₁ NO₅ C, 68.64, H, 5.77, N, 3.81 Found C, 68.21, H, 5.84, N,3.67. ¹ H NMR (CDCl₃) 7.53-7.67 (m, 10H), 5.67 (s, 1H), 5.16 (s, 2H),4.76 (broad s, 1H), 4.59 (d, J=15 Hz, 1H), 4.08 (d, J=10 Hz, 1H), 4.02(dd, J=11.4, 4 Hz, 1H), 3.46 (dd, J=15, 0.9 Hz, 1H), 3.40 (d, J=3 Hz,1H), 3.25 (d, J=3 Hz, 1H), 3.10 (dt, J=4, 10 Hz, 1H); ¹³ C NMR (CDCl₃)156.2, 137.8, 136.6, 129.7, 129.1, 128.9, 128.8, 128.5, 126.6, 102.8,73.0, 70.4, 68.0, 56.0, 54.7, 50.4 and 46.6; MS (CI, NH₃, m/e) 368(M+H).

EXAMPLE 5 Synthesis of2-azido-1,2,5-trideoxy-1,5-[{(phenylmethoxy)carbonyl}imino]-4,6-O-(R-phenylmethylene)-D-glucitol(7) and3-azido-1,3,5-trideoxy-1,5-[{(phenylmethoxy)carbonyl}imino]-4,6-O-(R-phenylmethylene)-D-altritol(6)

To a solution of epoxide 5 (4 g, 10.9 mmol) in 2-methoxyethanol (80 ml),sodium azide (3.5 g, 54.5 mmol) and ammonium chloride (2.33 g, 43.6mmol) were added. The reaction mixture was refluxed for 36 hr. Part ofthe solvent was removed under reduced pressure. The reaction mixture wasdiluted with ethyl acetate and washed with 1N HCl, water and brine. Theorganic layer was dried (MgSO₄), filtered and concentrated. The crudemixture was chromatographed (silica gel, hexane/ethyl acetate 8/2) togive pure 7 (1.95 g, 44%) and 6 (1.81 g, 41%). 6. DSC (mp) 253° C.; Analcalcd. for C₂₁ H₂₂ N₄₀₅ C, 61.46, H, 5.40, N, 13.65 Found C, 61.23, H,5.46, N, 13.39 7. Anal calcd. for C₂₁ H₂₂ N₄₀₅ C, 61.46, H, 5.40, N,13.65 Found C, 61.31, H, 5.56, N, 13.26.

EXAMPLE 6 Synthesis of2-azido-1,2,5-trideoxy-1,5-imino-4,6-O-(R-phenylmethylene)-D-glucitol(8)

Tho compound 7 (3.3 g, 8.05 mol) was added to previously preparedsolution of sodium hydroxide (4 g) in ethanol/water (1/1, 120 ml). Afterheating the mixture at 70° C. for 20 hr, the reaction was cooled andpart of the solvent was removed under reduced pressure. The mixture wasneutrallized with 1N HCl and extracted in methylene chloride. Theorganic layer was washed with water and brine. After drying (MgSO₄) andconcentration of the filterate, the crude product (3.01 g) waschromatographed (silica gel, ethyl acetae/i-propanol 98/2) to give pure8 (2.07 g, 93%). Anal calcd. for Cl₃ H₁₆ N₄₀₃ C, 56.51, H, 5.84, N,20.28 Found C, 56.56, H, 5.93, N, 20.15.

EXAMPLE 7 Synthesis of2-azido-1,5-(butylimino)-1,2,5-trideoxy-4,6-O-(R-phenylmethylene)-D-glucitol(9)

To a solution of 8 (3.1 g, 11.23 mmol) in methanol (120 ml), molecularsieves (4A°, 3.5 g) were added. After stirring for 5 min, butyraldehyde(1.86 ml, 20.8 mol), acetic acid (1.3 ml) and sodium cyanoborohydride(95%, 1.02 g, 15.4 mmol) were added. The reaction was stirred at 22° C.for 18 hr, filtered and the residue washed with more ethyl acetate. Thecombined organic fractions were concentrated. The residue wasredissolved in ethyl acetate and washed with aqueous potassiumcarbonate, water and brine. After drying (MgSO₄) and concentration, thecrude (4.08 g) was chromatographed (silica gel, hexane/ethyl acetate6/4) to give 9 (3.28 g, 88%) as white solid. DSC (mp) 115° C. (dec.);Anal calcd. for C₁₇ H₂₄ N₄₀₃ C, 61.43, H, 7.28, N, 16.85 Found C, 61.40,H, 7.34, N, 16.84.

EXAMPLE 8 Synthesis of2-azido-1,5-{(2-ethylbutyl)imino}-1,2,5-trideoxy-4,6-O-(R-phenylmethylene)-D-glucitol(10)

To a solution of 8 (1.07 g, 3.87 mmol) in methanol (35 ml), molecularsieves (4A°, 2.1 g) were added. After stirring for 5 min,2-ethylbutyraldehyde (1.04 ml, 7.74 mol), acetic acid (0.35 ml) andsodium cyanoborohydride (95%, 390 mg, 5.8 mmol) were added. The reactionwas stirred at 22° C. for 20 hr, filtered and the residue washed withmore ethyl acetate. The combined organic fractions were concentrated.The residue was redissolved in ethyl acetate and washed with aqueouspotassium carbonate, water and brine. After drying (MgSO₄) andconcentration, the crude (1.47 g) was chromatographed (silica gel,hexane/ethyl acetate 8/2) to give pure 10 (650 mg, 47%). Anal calcd. forC₁₉ H₂₈ N₄₀₃ C, 62.68, H, 7.86, N, 15.39 Found C, 62.72, H, 7.94, N,15.16.

EXAMPLE 9 Synthesis of2-azido-1,5-{(4,4,4-trifluorobutyl)imino}-1,2,5-trideoxy-4,6-O-(R-phenylmethylene)-D-glucitol(11)

To a solution of 8 (500 mg, 1.81 mmol) in dimethylformamide (10 ml),1-bromo-4,4,4-trifluorobutane (375 mg, 1.96 mmol) and potassiumcarbonate (150 mg, 1.08 mmol) were added. The reaction was immersed inan oil-bath at 60° C. and stirred for 60 hr. More1-bromo-4,4,4-trifluorobutane (375 mg, 1.96 mmol) was added and thereaction was heated at 60° C. for 24 hr. The solvent was removed underreduced pressure and the reaction mixture was neutralized with 1N HCl.The mixture was extracted in methylene chloride and the extract waswashed with aqueous potassium carbonate and brine. After drying (MgSO₄)and concentration, the crude (610 mg) was chromatographed (silica gel,hexane/ethyl acetate 6/4) to give pure 11 (510 mg, 73%) as thick liquid.¹ H NMR (CDCl₃) 7.49 (m, 2H), 7.39 (m, 3H), 5.49 (s, 1H), 4.34 (dd,J=11, 4 Hz, 1H), 3.63 (dd, J=11, 10 Hz, 1H), 3.45-3.60 (complex band,2H), 3.48 (t, J=9 Hz, 1H), 3.10 (d, J=2 Hz, 1H), 2.97 (dd, J=12, 5 Hz,1H), 2.55 (dt, J=13, 5 Hz, 1H), 2.36 (td, J=10, 4 Hz, 1H), 2.30 (dt,J=13, 7 Hz, 1H), 1.90-2.22 (complex band, 3H), 1.68 (d, J=7.5 Hz, 1H).

EXAMPLE 10 Synthesis of 2-azido-1,2,5-trideoxy-1,5-imino-D-glucitol (12)

A solution of 8 (1 g, 3.61 mmol) in trifluoroacetic acid/water (4/1, 15ml) was stirred at 22° C. for 18 hr. The solvent was removed underreduced pressure and the residue as thick yellow liquid was passedthrough an ion-exchange column [Amberlite, IRA-400 (OH)] prewashed withdistilled water until neutral. The basic fractions, as also followed byTLC (silica gel, ethyl acetate/methanol/ammonium hydroxide 50/50/2.5),were pooled and concentrated. The water in the fractions wasazeotropically removed with toluene and 12 (394 mg, 74%) was isolated asa white solid after crystallization from methanol/hexane, mp 142° C.(dec). Anal calcd. for C₆ H₁₂ N₄ O₃ 0.25H₂ O C, 39.86, H, 6.64, N, 27.75Found C, 39.91, H, 6.79, N, 27.59.

EXAMPLE 11 Synthesis of2-azido-1,5-(butylimino)-1,2,5-trideoxy-D-glucitol (13)

A solution of 9 (650 mg, 1.96 mmol) in trifluoroacetic acid/water (4/1,12 ml) was stirred at 22° C. for 8 hr. The solvent was removed underreduced pressure and the residue was passed through an ion-exchangecolumn [Amberlite, IRA-400 (OH)] prewashed with distilled water untilneutral. The basic fractions, as also followed by TLC (silica gel, ethylacetate/i-propanol/water/ammonium hydroxide 70/25/5/2), were pooled andconcentrated. The water in the fractions was azeotropically removed withtoluene to give 13 (330 mg) which was rechromatographed (silica gel,ethyl acetate/i-propanol/water/ammonium hydroxide 70/25/5/2) to givepure 13 (260 mg, 61%) as thick liquid. Anal calcd. for C₁₀ H₂₀ N₄ O₃0.2H₂ O C, 48.45, H, 8.29, N, 22.60 Found C, 48.49, H, 8.31, N, 22.41.

EXAMPLE 12 Synthesis of2-azido-1,5-{(2-ethylbutyl)imino}-1,2,5-trideoxy-D-glucitol (14)

A solution of 10 (250 mg, 0.69 mmol) in trifluoroacetic acid/water (4/1,7 ml) was stirred at 22° C. for 18 hr. The solvent was removed underreduced pressure and the residue was passed through an ion-exchangecolumn [Amberlite, IRA-400 (OH)] prewashed with distilled water untilneutral. The basic fractions, as also followed by TLC (silica gel, ethylacetate/i-propanol/water/ammonium hydroxide 70/25/5/2), were pooled andconcentrated. The water in the fractions was azeotropically removed withtoluene to give 14 (151 mg) which was rechromatographed (silica gel,ethyl acetate/i-propanol/water/ammonium hydroxide 70/25/5/2) to givepure 14 (52 mg, 27%). DSC (mp) 127° C. (dec). Anal calcd. for Cl₂ H₂₄ N₄O₃ C, 52.92, H, 8.88, N, 20.57 Found C, 52.67, H, 8.91, N, 20.48.

EXAMPLE 13 Synthesis of2-azido-1,5-{(4,4,4-trifluorobutyl)imino}-1,2,5-trideoxy-D-glucitol (15)

A solution of 11 (500 mg, 1.29 mmol) in trifluoroacetic acid/water (4/1,25 ml) was stirred at 22° C. for 24 hr. The solvent was removed underreduced pressure and the residue was passed through an ion-exchangecolumn Amberlite, IRA-400 (OH)] prewashed with distilled water untilneutral. The basic fractions, as also followed by TLC (silica gel, ethylacetate/i-propanol/water/ammonium hydroxide 70/25/5/2), were pooled andconcentrated. The water in the fractions was azeotropically removed withtoluene to give crude 15 (320 mg) which was rechromatographed (silicagel, ethyl acetate/i-propanol/water/ammonium hydroxide 70/25/5/2) togive pure 15 (272 mg, 70%) as white solid. DSC (mp) 107° C. (dec). Analcalcd. for C₁₀ H₁₇ N₄ O₃ F₃ C, 40.27, H, 5.75, N, 18.78 Found C, 40.12,H, 5.71, N, 18.60.

EXAMPLE 14 Synthesis of2-amino-1,5-(butylimino)-1,2,5-trideoxy-4,6-O-(R-phenylmethylene)-D-glucitol(17)

To a solution of 9 (700 mg, 2.11 m/mol) in methanol (70 ml) in a Parrhydrogenation flask, 10% Pd on C (70 mg) was added. The system wassealed, purged with nitrogen (5 times) and hydrogen (5 times) and thenpressurized to 5 psi hydrogen. After running the reaction on a shakerfor 3.5 hr, the system was vented, purged with nitrogen and filtered.The filtrate was concentrated and the crude (630 mg) was chromatographed(silica gel, methylene chloride/methanol/ammonium hydroxide 90/10/1) togive pure 17 (600 mg, 93%). DSC (mp) 125° C. Anal calcd. for C₁₇ H₂₆ N₂O₃ C, 66.64, H, 8.55, N, 9.14 Found C, 66.14, H, 8.56, N, 9.08.

EXAMPLE 15 Synthesis of2-amino-1,5-{(2-ethylbutyl)imino}-1,2,5-trideoxy-4,6-O-(R-phenylmethylene)-D-glucitol(18)

To a solution of 10 (350 mg, 0.97 mmol) in methanol (50 ml) in a Parrhydrogenation flask, 10% Pd on C (35 mg) was added. The system wassealed, purged with nitrogen (5 times) and hydrogen (5 times) and thenpressurized to 5 psi hydrogen. After running the reaction on a shakerfor 3 hr, the system was vented, purged with nitrogen and filtered. Thefiltrate was concentrated and the crude (320 mg) was chromatographed(silica gel, methylene chloride/methanol/ammonium hydroxide 90/10/1) togive pure 18 (240 mg, 78%). Anal calcd. for C₁₉ H₃₀ N₂ O₃ C, 68.23, H,9.04, N, 8.38 Found C, 68.87, H, 9.01, N, 7.48.

EXAMPLE 16 Synthesis of2-amino-1,5-{(4,4,4-trifluorobutyl)imino}-1,2,5-trideoxy-4,6-O-(R-phenylmethylene)-D-glucitol(19)

To a solution of 11 (1.4 g, 3.63 mmol) in methanol (25 ml) in a Parrhydrogenation flask, 10% Pd on C (140 mg) was added. The system wassealed, purged with nitrogen (5 times) and hydrogen (5 times) and thenpressurized to 5 psi hydrogen. After running the reaction on a shakerfor 21 hr, the system was vented, purged with nitrogen and filtered. Thefiltrate was concentrated and the crude (1.3 g) was chromatographed(silica gel, methylene chloride/methanol/ammonium hydroxide 90/10/1) togive pure 19 (1.15 g, 88%). Anal calcd. for C₁₇ H₂₃ N₂ O₃ F₃ 0.4H₂ O C,55.55, H, 6.53, N, 7.62 Found C, 55.55, H, 6.36, N, 7.59.

EXAMPLE 17 Synthesis of 2-amino-1,2,5-trideoxy-1,5-imino-D-glucitol (20)

To a solution of 12 (465 mg, 3.14 mmol) in methanol (50 ml) in a Parrhydrogenation flask, 10% Pd on C (50 mg) was added. The system wassealed, purged with nitrogen (5 times) and hydrogen (5 times) and thenpressurized to 5 psi hydrogen. After running the reaction on a shakerfor 3.5 hr, the system was vented, purged with nitrogen and filtered.The filtrate was concentrated to give pure 20 (365 mg, 91%). DSC (mp)184° C. Anal calcd. for C₆ H₁₄ N₂ O₃ 0.25H₂ O C, 43.23, H, 8.77, N,16.81 Found C, 43.42, H, 8.43, N, 16.47.

EXAMPLE 18 Synthesis of2-amino-1,5-(butylimino)-1,2,5-trideoxy-D-glucitol (21)

A solution of 17 (580 mg, 1.89 mmol) in trifluoroacetic acid/water (4/1,15 ml) was stirred at 22° C. for 24 hr. The solvent was removed underreduced pressure and the residue was passed through an ion-exchangecolumn [Amberlite, IRA-400 (OH)] prewashed with distilled water untilneutral. The basic fractions, as also followed by TLC (silica gel, ethylacetate/methanol/ammonium hydroxide 50/50/2.5), were pooled andconcentrated. The water in the fractions was azeotropically removed withtoluene to give crude 21 (410 mg) which was rechromatographed (silicagel, ethyl acetate/methanol/ammonium hydroxide 50/50/2.5) to give pure21 (302 mg, 73%). DSC (mp) 108° C. Anal calcd. for C₁₀ H₂₂ N₂ O₃ 0.3H₂ OC, 53.69, H, 10.18, N, 12.52 Found C, 53.63, H, 10.02, N, 12.34.

EXAMPLE 19 Synthesis of2-amino-1,5-{(2-ethylbutyl)imino}-1,2,5-trideoxy-D-glucitol (22)

A solution of 18 (140 mg, 0.42 mmol) in trifluoroacetic acid/water (4/1,10 ml) was stirred at 22° C. for 8 hr. The solvent was removed underreduced pressure and the residue was passed through an ion-exchangecolumn [Amberlite, IRA-400 (OH)] prewashed with distilled water untilneutral. The basic fractions, as also followed by TLC (silica gel, ethylacetate/i-propanol/ammonium hydroxide 50/50/2.5), were pooled andconcentrated. The water in the fractions was azeotropically removed withtoluene to give crude 22 (120 mg) which was rechromatographed (silicagel, ethyl acetate/i-propanol/ammonium hydroxide 50/50/2.5) to give pure22 (72 mg, 70%). DSC (mp) 130° C. Anal calcd. for Cl₂ H₂₆ N₂ O₃ 0.75 H₂O C, 55.46, H, 10.67, N, 10.78 Found C, 55.33, H, 10.05, N, 10.54.

EXAMPLE 20 Synthesis of2-amino-1,5-{(4,4,4-trifluorobutyl)imino}-1,2,5-trideoxy-D-glucitol (23)

A solution of 19 (400 mg, 1.1 mmol) in trifluoroacetic acid/water (4/1,10 ml) was stirred at 22° C. for 8 hr. The solvent was removed underreduced pressure and the residue was passed through an ion-exchangecolumn [Amberlite, IRA-400 (OH)] prewashed with distilled water untilneutral. The basic fractions, as also followed by TLC (silica gel, ethylacetate/i-propanol/ammonium hydroxide 50/50/2.5), were pooled andconcentrated. The water in the fractions was azeotropically removed withtoluene to give crude 23 (280 mg) which was rechromatographed (silicagel, ethyl acetate/i-propanol/ammonium hydroxide 50/50/2.5) to give pure23 (265 mg, 87%). Anal calcd. for C₁₀ H₁₉ N₂ O₃ F₃ 0.3H₂ O C, 43.26, H,7.11, N, 10.09 Found C, 43.23, H, 6.86, N, 9.59.

EXAMPLE 21 Synthesis of1,5-(butylimino)-1,2,5-trideoxy-2-(dimethylamino)-4,6-O-(R-phenylmethylene)-D-glucitol(24) and1,5-(butylimino)-1,2,5-trideoxy-2-(methylamino)-4,6-O-(R-phenylmethylene)-D-glucitol(25)

To a solution of 17 (792 mg, 2.59 mmol) in methanol (75 ml) in a Parrhydrogenation flask, 4% Pd on C (100 mg) and formaldehyde (0.23 ml )were added. The system was sealed, purged with nitrogen (5 times) andhydrogen (5 times) and then pressurized to 5 psi hydrogen. After runningthe reaction on a shaker for 21 hr, the system was vented, purged withnitrogen and filtered. The filtrate was concentrated and the crude (560mg) was chromatographed (silica gel, methylenechloride/methanol/ammonium hydroxide 90/10/1) to give 24 (310 mg, 36%)and 25 (372 mg, 45%). 24. ¹ H NMR (CDCl₃) 7.52 (m, 2H), 7.34 (m, 3H),5.52 (s, 1H), 4.37 (dd, J=11, 5 Hz, 1H), 3.83 (brs, 1H), 3.65 (dd, J=11,10 Hz, 1H), 3.60 (t, J=9 Hz, 1H), 3.54 (t, J=9 Hz, 1H), 2.93 (dd, J=11,4 Hz, 1H), 2.63 (ddd, J=11, 9, 4 Hz, 1H), 2.53 (dt, J=13, 8 Hz, 1H),2.35 (s, 6H), 2.22-2.37 (complex band, 2H), 2.14 (t, J=11 Hz, 1H), 1.42(m, 2H), 1.27 (m, 2H), 0.92 (t, J=7 Hz, 3H). 25. ¹ H NMR (CDCl₃) 7.49(m, 2H), 7.34 (m, 3H), 5.49 (s, 1H), 4.36 (dd, J=11, 4 Hz, 1H), 3.65(dd, J=11, 10 Hz, 1H), 3.48 (t, J=9 Hz, 1H), 3.36 (dd, J=10, 9 Hz, 1 H),3.25 (broad s, 1H), 3.05 (dd, J=11, 5 Hz, 1H), 2.57 (td, J=10, 5 Hz,1H), 2.51 (dt, J=13, 8 Hz, 1H), 2.39 (ddd, J=10, 9, 4 Hz, 1H), 2.37 (s,3H), 2.30 (ddd, J=13, 8, 6 Hz, 1H), 2.00 (t, J=11 Hz, 1H), 1.42 (m, 2H),1.26 (m, 2H), 0.92 (t, J=7 Hz, 3H).

EXAMPLE 221,5-(butylimino)-1,2,5-trideoxy-2-{(1-oxobutyl)amino}-4,6-O-(R-phenylmethylene)-D-glucitol,3-butanoate (26)

To a solution of 17 (650 mg 2.12 mmol) in pyridine (8 ml), butyricanhydride (2 ml) was added and the reaction mixture was stirred at roomtemperature. After stirring for 18 hr, the reaction mixture was pouredover ice and extracted with methylene chloride. The organic layer waswashed with water and brine. After drying over MgSO₄, the extract wasfiltered and the solvent removed under reduced pressure. The crudeproduct 26 (1.01 g) was used in the next step without furtherpurification. ¹ H NMR (CDCl₃) 7.45 (m, 2H), 7.36 (m, 3H), 5.86 (d, J=7.5Hz, 1H), 5.53 (s, 1H), 4.92 (t, J=10 Hz, 1H), 4.43 (dd, J=11, 5 Hz, 1H),4.16 (tdd, J=10, 7.5, 5 Hz, 1H), 3.74 (t, J=10 Hz, 1H), 3.73 (dd, J= 11,10 Hz, 1H), 3.25 (dd, J=12, 5 Hz, 1H), 2.55 (dt, J=13, 8 Hz, 1H), 2.46(td, J=10, 5 Hz, 1H), 2.35 (dt, J=15, 7.5 Hz, 1H), 2.29 (dt, J=15, 7.5Hz, 1H), 2.27 (dt, J=13, 7 Hz, 1H), 2.10 (t, J=7.5 Hz, 2H), 2.06 (dd,J=12, 10 Hz, 1H), 1.62 (m, 4H), 1.41 (m, 2H), 1.27 (m, 2H), 0.93 (t,J=7.5 Hz, 1H), 0.90 (t, J=7.5 Hz, 1H).

EXAMPLE 231,5-(butylimino)-1,2,5-trideoxy-2-{(1-oxobutyl)amino}-4,6-O-(R-phenylmethylene)-D-glucitol(27)

To a solution of 26 (900 mg, 2.01 mmol) in methanol (50 ml), saturatedaqueous potassium carbonate (30 ml) was added and the mixture wasstirred at room temperature for 4 hr. After neutrallizing with conc. HClto pH 7, methanol was removed under reduced pressure and the reactionmixture was extracted with ethyl acetate. The organic layer was washedwith water and brine, dried (MgSO₄) and filtered. The concentration ofthe extract gave 27 (720 mg, 90%). mp 172° C. (dec), Anal calcd. for C₂₁H₃₂ N₂ O₄, C, 66.99, H, 8.57, N, 7.44 Found C, 66.82, H, 8.68, N, 7.36.

EXAMPLE 24 Synthesis of1,5-(butylimino)-1,2,5-trideoxy-2-(dimethylamino)-D-glucitol (28)

A solution of 24 (580 mg, 1.74 mmol) in trifluoroacetic acid/water (4/1,10 ml) was stirred at 22° C. for 24 hr. The solvent was removed underreduced pressure and the residue was passed through an ion-exchangecolumn [Amberlite, IRA-400 (OH)] prewashed with distilled water untilneutral. The basic fractions, as also followed by TLC (silica gel, ethylacetate/i-propanol/ammonium hydroxide 50/50/2.5), were pooled andconcentrated. The water in the fractions was azeotropically removed withtoluene to give crude 28 (300 mg) which was rechromatographed (silicagel, ethyl acetate/i-propanol/ammonium hydroxide 50/50/2.5) to give pure28 (260 mg, 61%). DSC (mp) 111° C., Anal calcd. for C₁₂ H₂₆ N₂ O₃ 0.2 H₂O, C, 57.66, H, 10.65, N, 11.21 Found C, 57.88, H, 10.63, N, 11.23.

EXAMPLE 25 Synthesis of1,5-(butylimino)-1,2,5-trideoxy-2-(methylamino)-D-glucitol (29)

A solution of 25 (610 mg, 1.91 mmol) in trifluoroacetic acid/water (4/1,10 ml) was stirred at 22° C. for 24 hr. The solvent was removed underreduced pressure and the residue was passed through an ion-exchangecolumn [Amberlite, IRA-400 (OH)] prewashed with distilled water untilneutral. The basic fractions, as also followed by TLC (silica gel, ethylacetate/i-propanol/ammonium hydroxide 50/50/2.5), were pooled andconcentrated. The water in the fractions was azeotropically removed withtoluene to give crude 29 (480 mg) which was rechromatographed (silicagel, ethyl acetate/i-propanol/ammonium hydroxide 50/50/2.5) to give pure29 (310 mg, 70%). Anal calcd. for C₁₁ H₂₄ N₂ O₃ 0.4 H₂ O, C, 55.16, H,10.44, N, 11.70 Found C, 55.24, H, 10.57, N, 11.74.

EXAMPLE 261,5-(butylimino)-1,2,5-trideoxy-2-{(1-oxobutyl)amino}-D-glucitol (30)

A solution of 27 (250 mg, 0.66 mmol) in trifluoroacetic acid/water (4/1,10 ml) was stirred at 22° C. for 24 hr. The solvent was removed underreduced pressure and the residue was passed through an ion-exchangecolumn [Amberlite, IRA-400 (OH)] prewashed with distilled water untilneutral. The basic fractions, as also followed by TLC (silica gel, ethylacetate/i-propanol/water/ammonium hydroxide 70/25/5/2), were pooled andconcentrated. The water in the fractions was azeotropically removed withtoluene to give crude 30 (180 mg) which was rechromatographed (silicagel, ethyl acetate/i-propanol/water/ammonium hydroxide 70/25/5/2) togive pure 30 (165 mg, 86%). DSC (mp) 203° C., Anal calcd. for C₁₄ H₂₈ N₂O₄ 0.5 H₂ O, C, 56.54, H, 9.83, N, 9.42 Found C, 56.32, H, 9.50, N,9.26.

EXAMPLE 271,5-(butylimino)-1,2,5-trideoxy-2-{(1-oxobutyl)amino}-D-glucitol,tributanoate (31)

To a solution of 30 (100 mg 0.34 mmol) in pyridine (8 ml), butyricanhydride (2 ml) was added and the reaction mixture was stirred at roomtemperature. After stirring for 40 hr, the reaction mixture was pouredover ice and extracted with methylene chloride. The organic layer waswashed with water and brine. After drying over MgSO₄, the extract wasfiltered and the solvent removed under reduced pressure. The crudeproduct (280 mg) was chromatographed (silica gel, hexane/ethyl acetate6/4) to give pure 31 (98 mg, 57%). DSC (mp) 84° C. (dec), Anal calcd.for C₂₆ H₄₆ N₂ O₇, C, 62.63, H, 9.30, N, 5.62 Found C, 62.17, H, 9.30,N, 5.32.

EXAMPLE 28 Synthesis of phenylmethyl8β-azidohexahydro-7-oxo-2R-2α-phenyl-5H-4aα,8αβ-1,3-dioxino[5,4-b]pyridine-5-carboxylate (32)

To a cold solution of dimethyl sulfoxide (5.6 ml, 78 mmol) in methylenechloride (50 ml) at -70° C., trifluoroacetic anhydride (8.32 ml, 59mmol) in methylene chloride (50 ml) was added over 20 min. Afterstirring for 15 min, a solution of 6 (16 g, 39 mmol) in methylenechloride (150 ml) was added over 30 min. at -70° C. The temperature ofreaction mixture was allowed to rise to -30° C. over 4 hr and then thereaction was stirred at -30° C. for 1 hr. After recooling to -70° C.,triethylamine (15 ml, 107 mmol) was added and the reaction was warmed to22° C. in about an 1 hr and stirred at 22° C. for about 8 hr. Thereaction was diluted with methylene chloride and washed with water andbrine. After drying (MgSO₄), filteration and concentration, the crude(17.8 g) was chromatographed (silica gel, hexane/ethyl acetate 1/1) togive pure 32 (13.9 g, 86%). ¹ H NMR (CDCl₃) 7.30-7.49 (complex band,10H), 5.71 (s, 1H), 5.13 (s, 2H), 4.85 (d, J=11 Hz, 1H), 4.61 (dd, J=11,4 Hz, 1H), 4.33 (dd, J=11, 10 Hz, 1H), 4.30 (d, J=18 Hz, 1H), 4.20 (d,J=18 Hz, 1H), 4.11 (dd, J=11, 10, 1H), 3.85 (dt, J=10, 4Hz, 1H).

EXAMPLE 293-azido-1,3,5-trideoxy-1,5-[{(phenylmethoxy)carbonyl}imino]-4,6-O-(R-phenylmethylene)-D-glucitol(33) and3-azido-1,3,5-trideoxy-1,5-[{(phenylmethoxy)carbonyl}imino]-4,6-O-(R-phenylmethylene)-D-mannitol(34)

To a cold solution of 32 (2.5 g, 6.12 mmol) in THF (100 ml) at -78° C.,diisobutylaluminum hydride (9.25 ml, 1M solution in toluene, 9.25 mmol)was added over 10 min. After stirring at -78° C. for 4 hr, methanol (2.5ml) was added. The reaction was stirred for 10 min, the cold bath wasremoved and the reaction allowed to rise to 22° C. and stirred for 30min. After quenching with 0.5N HCl (10 ml), the reaction was dilutedwith ethyl acetate and washed with water and brine. The organic extractwas dried (MgSO₄), filtered and concentrated to give crude mixture (2.23g) as thick orange liquid. Chromatographic purification (silica gel,hexane/ethyl acetate 1/1) gave 33 (1.57 g, 63%) and 34 (231 mg, 9%). 33.Anal calcd. for C₂₁ H₂₂ N₄ O₅, C, 61.46, H, 5.40, N, 13.65 Found C,61.62, H, 5.53, N, 12.48. 34. Anal calcd. for C₂₁ H₂₂ N₄ O₅, C, 61.46,H, 5.40, N, 13.65 Found C, 61.37, H, 5.43, N, 13.39.

EXAMPLE 303-azido-1,3,5-trideoxy-1,5-imino-4,6-O-(R-phenylmethylene)-D-glucitol(35)

Tho compound 33 (1.8 g, 4.39 mol) was added to previously preparedsolution of sodium hydroxide (2 g) in ethanol/water (1/1, 60 ml). Afterheating the mixture at 75°-80° C. for 20 hr, the reaction was cooled andpart of the solvent was removed under reduced pressure. The mixture wasneutrallized with 1N HCl and extracted in methylene chloride. Theorganic layer was washed with water and brine. After drying (MgSO₄) andconcentration of filterate, the crude product (3.01 g) waschromatographed (silica gel, methylene chloride/methanol/ammoniumhydroxide 90/10/1) to give pure 35 (1.1 g, 91%). DSC (mp) 192° C., Analcalcd. for Cl₃ H₁₆ N₄ O₃, C, 56.51, H, 5.84, N, 20.28 Found C, 56.26, H,5.90, N, 20.08.

EXAMPLE 31 3-amino-1,3,5-trideoxy-1,5-imino-4,6-O-(R-phenylmethylene)-D-glucitol(36)

To a solution of 35 (700 mg, 2.54 mmol) in methanol (50 ml) in a Parrhydrogenation flask, 4% Pd on C (150 mg) was added. The system wassealed, purged with nitrogen (5 times) and hydrogen (5 times) and thenpressurized to 5 psi hydrogen. After running the reaction on a shakerfor 10 hr, the system was vented, purged with nitrogen and filtered. Thefiltrate was concentrated and the crude (700 mg) was chromatographed(silica gel, methylene chloride/methanol/ammonium hydroxide 90/10/1) togive pure 36 (590 mg, 93%). Anal calcd. for Cl₃ H₁₈ N₂ O₃ 0.25H₂ O, C,61.28, H, 7.32, N, 10.99 Found C, 61.27, H; 7.29, N, 10.72.

EXAMPLE 32 3-amino-1,3,5-trideoxy-1,5-imino-D-glucitol (37)

A solution of 3{(480 mg, 1.92 mmol) in trifluoroacetic acid/water (4/1,8 ml) was stirred at 22° C. for 24 hr. The solvent was removed underreduced pressure and the residue was passed through an ion-exchangecolumn [Amberlite, IRA-400 (OH)] prewashed with distilled water untilneutral. The basic fractions, as also followed by TLC (silica gel, ethylacetate/methanol/ammonium hydroxide 25/75/3), were pooled andconcentrated. The water in the fractions was azeotropically removed withtoluene to give crude 37 which was rechromatographed (silica gel, ethylacetate/methanol/ammonium hydroxide 25/75/3) to give pure 37 (135 mg,32%). DSC (mp) 191° C., Anal calcd. for C₆ H₁₄ N₂ O₃ 0.25H2O, C, 43.23,H, 8.77, N, 16.81 Found C, 43.66, H, 8.61, N, 16.19.

EXAMPLE 33 Synthesis of2-azido-1,2,5-trideoxy-1,5-[{(phenylmethoxy)carbonyl}imino]-4,6-O-(R-phenylmethylene)-D-glucitol,methanesulfonate (38)

To a solution of 7 (3.8 g, 9.27 mmol) in pyridine (40 ml),methanesulfonyl chloride (860 μl, 11.11 mmol) was injected over 10 min.After stirring at 22° C. for 20 hr, the reaction contents were pouredover ice and extracted in ethyl acetate (2×700). The combined organicextracts were washed with saturated aqueous potassium carbonate, waterand brine. After drying (MgSO₄), filteration and concentration, thecrude (6.45 g) was chromatographed (silica gel, hexane/ethyl acetate6/4) to give pure 38 (4.3 g, 95%) as white solid. DSC (mp) 222° C., Analcalcd. for C₂₂ H₂₄ N₄ O₇ S 1H2O, C, 52.17, H, 5.17, N, 11.06 Found C,52.29, H, 4.81, N, 10.87.

EXAMPLE 34 Synthesis of2-azido-1,2,5-trideoxy-1,5-[{(phenylmethoxy)carbonyl}imino]-4,6-O-(R-phenylmethylene)-D-allitol,acetate (39)

A mixture of 38 (2.3 g, 4.7 mmol), cesium acetate (9 g, 47 mmol),18-crown-6 (1.16 g, 4.7 mmol) in toluene (50 ml) was refluxed for 72 hr.The reaction was cooled, filtered and the residue washed with moretoluene. The combined organic fractions were concentrated and the crude(3.36 g) was chromatographed (silica gel, hexane/ethyl acetate 7/3) togive pure 39 (1.1 g, 52%) as white solid in addition to the startingmaterial 38 (0.31 g, 14%). 39. ¹ H NMR (CDCl₃) 7.31-7.45 (complex band,10H), 5.74 (td, J=3, 1 Hz, 1H), 5.56 (s, 1H), 5.15 (d, J=12 Hz, 1H).5.11 (d, J=12 Hz, 1H), 4.84 (dd, J=12, 5 Hz, 1H), 4.48 (dd, J=12, 10 Hz,1H), 4.32 (ddd, J=13, 5, 1 Hz, 1H), 3.79 (dd, J=10, 3 Hz, 1H), 3.61 (td,J=10, 5 Hz, 1H), 3.52 (ddd, J=11, 5, 3 Hz, 1H), 3.16 (dd, J=13, 11 Hz,1H), 2.17 (s, 3H).

EXAMPLE 35 Synthesis of2-azido-1,2,5-trideoxy-1,5-[{(phenylmethoxy)carbonyl}imino]-4,6-O-(R-phenylmethylene)-D-allitol(40A) and2-azido-1,2,5-trideoxy-1,5-{(methoxycarbonyl)imino}-4,6-O-(R-phenylmethylene)-D-allitol(40B)

A mixture of 39 (970 mg, 2.15 m/mol) and sodium methoxide (400 mg, 7.4mmol) in methanol (50 ml) was refluxed for 18 hr. The reaction wascooled, neutrallized with 1N HCl and the solvent removed under reducedpressure. The residue was suspended in ethyl acetate and washed withsaturated aqueous potassium carbonate, water and brine. The combinedorganic extracts were concentrated and the crude (1.02 g)chromatographed (silica gel, hexane/ethyl acetate 7/3) to give 40A (550rag, 57%) and 40B (270 mg, 35%). 40A. ¹ H NMR (CDCl₃) 7.45 (m, 2H), 7.34(m, 8H), 5.55 (s, 1H), 5.10 (d, J=12 Hz, 1H). 5.07 (d, J=12 Hz, 1H),4.79 (dd, J=12, 5 Hz, 1H), 4.45 (dd, J=12, 10 Hz, 1H), 4.22 (broad s,1H), 4.17 (m, 1H), 3.62 (td, J=10, 5 Hz, 1H), 3.54 (dt, J=10, 2 Hz, 1H),3.24 (m, 1H), 3.21 (m, 1H), 2.87 (s, 1H). 40B. 1H NMR (CDCl₃) 7.47 (m,2H), 7.37 (m, 3H), 5.59 (s, 1H), 4.81 (dd, J=12, 4 Hz, 1H), 4.48 (dd,J=12, 9 Hz, 1H), 4.27 (broad s, 1H), 4.12 (dd, J=12, 2 Hz, 1H), 3.67 (s,3H), 3.65 (m, 1H), 3.60 (m, 1H), 3.30 (m, 1H), 3.23 (m, 1H), 2.82 (broads, 1H).

EXAMPLE 36 Synthesis of2-azido-1,2,5-trideoxy-1,5-[{(phenylmethoxy)carbonyl}imino]-4,6-O-(R-phenylmethylene)-D-allitol,methanesulfonate (41A)

To a solution of 40A (550 mg, 1.34 mmol) in pyridine (10 ml),methanesulfonyl chloride (140 μl, 1.74 mmol) was injected over 10 min.After stirring at 22° C. for 60 hr, the reaction contents were pouredover ice and extracted in ethyl acetate. The combined organic extractswere washed with saturated aqueous potassium carbonate, water and brine.After drying (MgSO₄), filteration and concentration, the productobtained 41A (603 mg, 92%) was used in the next step without furtherpurification. ¹ NMR (CDCl₃) 7.44 (m, 2H), 7.35 (m, 8H), 5.58 (s, 1H),5.15 (broad t, J=2.5 Hz, 1H), 5.11 (s, 2H), 4.87 (dd, J=12, 5 Hz, 1H),4.45 (dd, J=12, 10 Hz, 1H), 4.31 (dd, J=13, 5 Hz, 1H), 3.80 (dd, J= 10,2 Hz, 1H), 3.60 (ddd, J=12, 5, 3 Hz, 1H), 3.56 (td, J=10, 5 Hz, 1H),3.10 (dd, J=13, 12 Hz, 1H), 2.92 (s, 3H).

EXAMPLE 37 Synthesis of2-azido-1,2,5-trideoxy-1,5-{(methoxycarbonyl)imino}-4,6-O-(R-phenylmethylene)-D-allitol,methanesulfonate (41B)

To a solution of 40B (217 mg, 0.65 mmol) in pyridine (5 ml),methanesulfonyl chloride (65 μl, 0.84 mmol) was injected over 10 min.After stirring at 22° C. for 30 hr, the reaction contents were pouredover ice and extracted in ethyl acetate. The combined organic extractswere washed with saturated aqueous potassium carbonate, water and brine.After drying (MgSO₄), filteration and concentration, the productobtained 41B (320 mg, 92%) was used in the next step without furtherpurification. ¹ H NMR (CDCl₃) 7.46 (m, 2H), 7.35 (m, 3H), 5.61 (s, 1H),5.18 (broad t, J=2.5 Hz, 1H), 4.88 (dd, J=12, 5 Hz, 1H), 4.48 (dd, J=12,10 Hz, 1H), 4.28 (dd, J=13, 5 Hz, 1H), 3.83 (dd, J=10, 2 Hz, 1H), 3.70(s, 3H), 3.66 (ddd, J=12, 5, 3 Hz, 1H), 3.58 (td, J=10, 5 Hz, 1H), 3.12(dd, J=13, 12 Hz, 1H), 2.95 (s, 3H).

EXAMPLE 38 Synthesis of2,3-diazido-1,2,3,5-tetradeoxy-1,5-{(methoxycarbonyl)imino}-4,6-O-(R-phenylmethylene)-D-glucitol(42B)

To a solution of 41B (320 mg, 0.77 mmol) in dimethylformamide (10 ml),sodium azide (252 mg, 3.88 mmol) was added. The reaction mixture washeated at 100°-10° C. for 30 hr. Part of the solvent was removed underreduced pressure. The reaction mixture was diluted with ethyl acetateand washed with aqueous potassium carbonate, water and brine. Theorganic layer was dried (MgSO₄), filtered and concentrated. The crude42B (190 mg, 69%) was used in the next step without furtherpurification. ¹ H NMR (CDCl₃) 7.50 (m, 2H), 7.37 (m, 3H), 5.62 (s, 1H),4.79 (dd, J=12, 5 Hz, 1H), 4.45 (dd, J=12, 10 Hz, 1H), 4.28 (dd, J=14, 5Hz, 1H), 3.68 (s, 3H), 3.67 (t, J=10 Hz, 1H), 3.50 (t, J=10 Hz, 1H),3.30 (ddd, J=11, 10, 5 Hz, 1H), 3.20 (td, J=10, 5 Hz, 1H), 2.64 (dd,J=14, 11 Hz, 1H).

EXAMPLE 39 Synthesis of2,3-diazido-1,2,3,5-tetradeoxy-1,5-[{(phenylmethoxy)carbonyl}imino]-4,6-O-(R-phenylmethylene)-D-glucitol(42A)

To a solution of 41A (600 mg, 1.23 mmol) in dimethylformamide (10 ml),sodium azide (400 mg, 6.15 mmol) was added. The reaction mixture washeated at 100°-10° C. for 72 hr. Part of the solvent was removed underreduced pressure. The reaction mixture was diluted with ethyl acetateand washed with aqueous potassium carbonate, water and brine. Theorganic layer was dried (MgSO₄), filtered and concentrated. The crudemixture (760 mg) consisting of 42A and 43 was hydrolyzed to 43 withoutpurification.

EXAMPLE 40 Synthesis of2,3-diazido-1,2,3,5-tetradeoxy-1,5-imino-4,6-O-(R-phenylmethylene)-D-glucitol(43)

The mixture of 42A, 42B & 43 (920 mg) obtained in the above steps wasadded to previously prepared solution of sodium hydroxide (2 g) inethanol/water (1/1, 60 ml). After refluxing the mixture for 20 hr, thereaction was cooled and part of the solvent was removed under reducedpressure. The mixture was neutrallized with 1N HCl and extracted inethyl acetate. The organic layer was washed with water and brine. Afterdrying (MgSO₄) and concentration of filterate, the crude product (280mg) was chromatographed (silica gel, methylene chloride/ethanol 98/2) togive pure 43. (360 mg, 68% in two steps). ¹ H NMR (CDCl₃) 7.56 (m, 2H),7.43 (m, 3H), 5.61 (s, 1H), 4.23 (dd, J=11, 5 Hz, 1H), 3.58 (dd, J=11,10 Hz, 1H), 3.53 (dd, J=10, 9 Hz, 1H), 3.42 (dd, J=10, 9 HZ, 1H), 3.29(td, J=10, 5 Hz, 1H), 3.21 (dd, J=12, 5 Hz, 1H), 2.71 (td, J=10, 5 Hz,1H), 2.54 (dd, J=12, 10 Hz, 1H), 1.15 (broad s, 1H).

EXAMPLE 412,3-diazido-1,5-(butylimino)-1,2,3,5-tetradeoxy-4,6-O-(R-phenylmethylene)-D-glucitol(44)

To a solution of 43 (360 mg, 1.19 mmol) in methanol (10 ml), molecularsieves (4A°, 0.7 g) were added. After stirring for 5 min, butyraldehyde(0.22 ml, 2.4 mol), acetic acid (0.2 ml ) and sodium cyanoborohydride(95%, 111 mg, 1.78 mmol) were added. The reaction was stirred at 22° C.for 18 hr, filtered and the residue washed with more methanol. Thecombined organic fractions were concentrated. The residue wasredissolved in ethyl acetate and washed with aqueous potassiumcarbonate, water and brine. After drying (MgSO₄) and concentration, thecrude (0.47 g) was chromatographed (silica gel, hexane/ethyl acetate8/2) to give pure 44 (410 mg, 94%). ¹ H NMR (CDCl₃ ) 7.50 (m, 2H), 7.37(m, 3H), 5.58 (s, 1H), 4.43 (dd, J=11, 5 Hz, 1H), 3.68 (dd, J=11, 10 Hz,1H), 3.59 (t, J=9 Hz, 1H), 3.45 (dd, J=10, 9 Hz, 1H), 3.38 (td, J=10, 5Hz, 1H), 3.07 (dd, J=12, 5 Hz, 1H), 2.53 (dt, J=13, 8 Hz, 1H), 2.41(ddd, J=10, 9, 5 Hz, 1H), 2.30 (dt, J=13, 7 Hz, 1H), 2.17 (dd, J=12, 10Hz, 1H), 1.39 (m, 2H), 1.28 (m, 2H), 0.92 (t, J=7 Hz, 3H) .

EXAMPLE 422,3-diamino-1,5-(butylimino)-1,2,3,5-tetradeoxy-4,6-O-(R-phenylmethylene)-D-glucitol(45)

To a solution of 44 (385 mg, 1.08 mmol) in methanol (25 ml) in a Parrhydrogenation flask, 10% Pd on C (60 mg) was added. The system wassealed, purged with nitrogen (5 times) and hydrogen (5 times) and thenpressurized to 5 psi hydrogen. After running the reaction on a shakerfor 3.5 hr, the system was vented, purged with nitrogen and filtered.The filtrate was concentrated and the crude (320 mg) was chromatographed(silica gel, ethyl acetate/methanol/ammonium hydroxide 50/50/2.5) togive 45 (240 mg, 73%). Anal calcd. for C₁₇ H₂₇ N₃ O₂ 0.25H₂ O, C, 65.88,H, 8.94, N, 13.56 Found C, 65.53, H, 8.99, N, 13.28.

EXAMPLE 432,3-diamino-1,5-(butylimino)-1,2,3,5-tetradeoxy-D-glucitol(46)

A solution of 45 (235 mg, 0.77 mmol) in trifluoroacetic acid/water (4/1,10 ml) was stirred at 22° C. for 18 hr. The solvent was removed underreduced pressure and the residue was passed through an ion-exchangecolumn [Amberlite, IRA-400 (OH)] prewashed with distilled water untilneutral. The basic fractions, as also followed by TLC (silica gel, ethylacetate/methanol/ammonium hydroxide 25/75/3), were pooled andconcentrated. The water in the fractions was azeotropically removed withtoluene to give crude 46 (152 mg) which was rechromatographed (silicagel, ethyl acetate/methanol/ammonium hydroxide 25/75/3) to give pure 46(72 mg, 43%). Anal calcd. for C₁₀ H₂₃ N₃ O₂, C, 55.27, H, 10.67, N,19.34 Found C, 54.86, H, 10.78, N, 19.00.

EXAMPLE 44 Synthesis of1,3,5-trideoxy-3-[{2-(dimethylamino)ethyl}amino]-1,5-[{(phenylmethoxy)carbonyl}imino]-4,6-O-(R-phenylmethylene)-D-altritol(47)

A solution of epoxide 5 (734 mg, 2 mmol) in N,N-dimethylaminoethylamine(7 ml) was heated at 100° C. for 24 hr. Part of the solvent was removedunder reduced pressure and the crude residue was chromatographed (silicagel, methylene chloride/methanol/ammonium hydroxide 90/10/1) to givepure 47 (700 mg, 76%) as an oil. Anal calcd. for C₂₅ H₃₃ N₃ O₅, C,65.91, H, 7.30, N, 9.22 Found C, 65.65, H, 7.45, N, 9.02.

EXAMPLE 45 Synthesis of3-(butylamino)-1,3,5-trideoxy-1,5-[{(phenylmethoxy)carbonyl}imino]-4,6-O-(R-phenylmethylene)-D-altritol(48)

A solution of epoxide 5 (200 mg, 0.55 mmol) in butylamine (4 ml) wasrefluxed for 24 hr. Part of the solvent was removed under reducedpressure and the crude residue was chromatographed (silica gel,hexane/ethyl acetate 70/30) to give pure 48 (117 mg, 70%). mp 104°-6°C., Anal calcd. for C₂₅ H₃₂ N₂ O₅, C, 68.16, H, 7.32, N, 6.36 Found C,68.04, H, 7.39, N, 6.34.

EXAMPLE 46 Synthesis of1,3,5-trideoxy-3-[{2-(dimethylamino)ethyl}amino]-1,5-imino-4,6-O-(R-phenylmethylene)-D-altritol(49)

To a solution of 47 (1.78 g, 3.9 mmol) in ethanol (35 ml) in a Parrhydrogenation flask, 4% Pd on C (250 mg) was added. The system wassealed, purged with nitrogen (5 times) and hydrogen (5 times) and thenpressurized to 5 psi hydrogen. After running the reaction on a shakerfor 5 hr, the system was vented, purged with nitrogen and filtered. Thefiltrate was concentrated and the crude was crystallized fromcyclohexane to give 49 (1.14 g, 91%). mp 100°-2° C., Anal calcd. for C₁₇H₂₇ N₃ O₃, C, 63.53, H, 8.47, N, 13.07 Found C, 63.28, H, 8.59, N,12.85.

EXAMPLE 47 Synthesis of3-[{2-(dimethylamino)ethyl}amino]-1,3,5-trideoxy-1,5-imino-D-altritol(50)

A solution of 49 (600 mg, 1.8 mmol) in trifluoroacetic acid/water (4/1,6 ml) was stirred at 25° C. for 25 hr. The solvent was removed underreduced pressure and the residue was passed through an ion-exchangecolumn [Amberlite, IRA-400 (OH)] prewashed with distilled water untilneutral. The basic fractions, as also followed by TLC (silica gel, ethylacetate/methanol/ammonium hydroxide 25/75/3), were pooled andconcentrated. The water in the fractions was azeotropically removed withtoluene to give 50 (250 mg, 72%) which was recrystallized from methanol.mp 120°-22° C., Anal calcd. for C₁₀ H₂₃ N₃ O₃, C, 51.47, H, 9.93, N,18.00 Found C, 51.61, H, 9.72, N, 17.81.

EXAMPLE 48

Various illustrative compounds synthesized above were tested forinhibition of visna virus in vitro in a plaque reduction assay (MethodA) or for inhibition of HIV-1 in a test which measured reduction ofcytopathogenic effect in virus-infected synctium-sensitiveLeu-3a-positive CEM cells grown in tissue culture (Method B) as follows:

Method A Cell and Virus Propagation

Sheep choroid plexus (SCP) cells were obtained from American TypeCulture Collection (ATCC) catalogue number CRL 1700 and were routinelypassaged in vitro in Dulbecco's Modified Eagles (DME) mediumsupplemented with 20% fetal bovine serum (FBS). SCP cells were passagedonce per week at a 1:2 or 1:3 split ratio. Visna was titrated by plaqueassay in six-well plates. Virus pools were stored at -70° C.

Plaque Reduction Assay

SCP cells were cultured in 6-well plates to confluence. Wells werewashed two times with serum free Minimal Essential Medium (MEM) toremove FBS. 0.2 ml of virus was added per well in MEM supplemented with4 mM glutamine and gentamycin. After 1 hour adsorption, the virus wasaspirated from each well. The appropriate concentration of each compoundin 5 ml of Medium 199 (M-199) supplemented with 2% lamb serum, 4 mMglutamine, 0.5% agarose and gentamycin was added to each well. Cultureswere incubated at 37° C. in a humidified 5% CO₂ incubator for 3-4 weeks.To terminate the test; cultures were fixed in 10% formalin, the agarremoved, the monolayers stained with 1% crystal violet and plaquescounted. Each compound concentration was run in triplicate. Controlwells (without virus) were observed for toxicity of compounds at thetermination of each test and graded morphologically from 0 to 4. 0 is notoxicity observed while 4 is total lysing of the cell monolayer.

96 Well Plate Assay

The 96 well plate assay was performed similarly to the plaque assayabove with modifications. SCP cells were seeded at 1×10⁴ cells per wellin 0.1 ml DME medium. When confluent, the wells were washed with serumfree MEM and 25 μl of virus added in M-199 supplemented with 2% lambserum. After 1 hour, 75 μL of medium containing test compound was addedto each well containing virus. After 2-3 weeks incubation the cytopathiceffect of the virus was determined by staining with a vital stain. Cellviability was measured by determining stain density using a 96 wellplate reader.

Control wells without virus were completed to determine the toxicity ofcompounds.

Method B

Tissue culture plates were incubated at 37° C. in a humidified, 5% CO₂atmosphere and observed microscopically for toxicity and/orcytopathogenic effect (CPE). At 1 hour prior to infection each testarticle was prepared from the frozen stock, and a 20 μl volume of eachdilution (prepared as a 10×concentration) was added to the appropriatewells of both infected and uninfected cells.

On the 9th day post-infection, the cells in each well were resuspendedand a 100 μl sample of each cell suspension was removed for use in anMTT assay. A 20 μl volume of a 5 mg/ml solution of3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) wasadded to each 100 μl cell suspension, and the cells were incubated at37° C. in 5% CO₂ for 4 hours. During this incubation MTT ismetabolically reduced by living cells, resulting in the production of acolored formazan product. A 100 μl volume of a solution of 10% sodiumdodecyl sulfate in 0.01N hydrochloric acid was added to each sample, andthe samples were incubated overnight. The absorbance at 590 nm wasdetermined for each sample using a Molecular Devices V_(max) microplatereader. This assay detects drug-induced suppression of viral CPE, aswell as drug cytotoxicity, by measuring the generation of MTT-formazanby surviving cells.

Assays were done in 96-well tissue culture plates. CEM cells weretreated with polybrene at a concentration of 2 μg/ml, and an 80 μlvolume of cells (1×10⁴ cells) was dispensed into each well. A 100 μlvolume of each test article dilution (prepared as a 2×concentration) wasadded to 5 wells of cells, and the cells were incubated at 37° C. for 1hour. A frozen culture of HIV-1, strain HTVL-III_(B), was diluted inculture medium to a concentration of 5×10⁴ TCID₅₀ per ml, and a 20 μlvolume (containing 10³ TCID₅₀ of virus) was added to 3 of the wells foreach test article concentration. This resulted in a multiplicity ofinfection of 0.1 for the HIV-1 infected samples. A 20 μl volume ofnormal culture medium was added to the remaining wells to allowevaluation of cytotoxicity. Each plate contained 6 wells of untreated,uninfected, cell control samples and 6 wells of untreated, infected,virus control samples.

Tables 2-6, below, set forth the results of the assay for illustrativecompounds prepared in the foregoing Examples in Method A: These resultsare stated in terms of % Plaque Reduction (mM concentration).

                  TABLE 2                                                         ______________________________________                                        Anti-Viral Activity of 2-Azido Analogs                                         ##STR10##                                                                                            Visna                                                 Compound   R            % Plaque Redn (Conc)                                  ______________________________________                                        12         H            76 (0.1 mM)                                           13         n-Bu         78 (0.1 mM)                                           14         CH.sub.2 CH(Et).sub.2                                                                      67 (0.1 mM)                                           15         (CH.sub.2).sub.3 CF.sub.3                                                                  41 (1 mM)                                             ______________________________________                                    

                  TABLE 3                                                         ______________________________________                                        Anti-Viral Activity of 2-Amino Analogs                                         ##STR11##                                                                                            Visna                                                 Compound   R            % Plaque Redn (Conc)                                  ______________________________________                                        20         H            91 (0.1 mM)                                           21         n-Bu         66 (0.1 mM)                                           22         CH.sub.2 CH(Et).sub.2                                                                      >1 mm                                                 23         (CH.sub.2).sub.3 CF.sub.3                                                                  9 (1 mM)                                              ______________________________________                                    

                  TABLE 4                                                         ______________________________________                                        Anti-Viral Activity of 2-Substituted Analogs                                   ##STR12##                                                                                         Visna                                                    Compound   X.sub.1   % Plaque Redn (Conc)                                     ______________________________________                                        13         N.sub.3   78 (0.1 mM)                                              21         NH.sub.2  66 (0.1 mM)                                              28         NMe.sub.2 41 (0.1 mM)                                              29         NHMe      25 (0.1 mM)                                              30         NHCOPr    20 (0.1 mM)                                              31         NHCOPr    29 (0.1 mM)                                                         (Per Butyr)                                                        ______________________________________                                    

                  TABLE 5                                                         ______________________________________                                        Anti-Viral Activity of C-2 & C-3 Substituted Analogs                           ##STR13##                                                                                             Visna                                                Compound  X.sub.1, X.sub.2                                                                             % Plaque Redn (Conc)                                 ______________________________________                                        37 (R = H)                                                                              X.sub.1 = H, X.sub.2 = NH.sub.2                                                              30 (1 mM)                                            46 (R = Bu)                                                                             X.sub.1 = X.sub.2 = NH.sub.2                                                                  3 (1 mM)                                            ______________________________________                                    

                  TABLE 6                                                         ______________________________________                                        Anti-Viral Activity of 3-amino Analog                                          ##STR14##                                                                                Visna                                                             Compound    % Plaque Redn (Conc)                                              ______________________________________                                        50          38 (1 mM)                                                         ______________________________________                                    

Compound (50) also effectively inhibited both α- and β-glucosidaseenzymes 22% at 1 mM concentration as determined in conventional assaysfor these enzymes described in U.S. Pat. No. 4,973,602.

The antiviral agents described herein can be used for administration toa mammalian host infected with a virus, e.g. visna virus or in vitro tothe human immunodeficiency virus, by conventional means, preferably informulations with pharmaceutically acceptable diluents and carriers.These agents can be used in the free amine form or in their salt form.Pharmaceutically acceptable salt derivatives are illustrated, forexample, by the HCl salt. The amount of the active agent to beadministered must be an effective amount, that is, an amount which ismedically beneficial but does not present toxic effects which overweighthe advantages which accompany its use. It would be expected that theadult human dosage would normally range upward from about one milligramof the active compound. The preferable route of administration is orallyin the form of capsules, tablets, syrups, elixirs and the like, althoughparenteral administration also can be used. Suitable formulations of theactive compound in pharmaceutically acceptable diluents and carriers intherapeutic dosage form can be prepared by reference to general texts inthe field such as, for example, Remington's Pharmaceutical Sciences, Ed.Arthur Osol, 16th ed., 1980, Mack Publishing Co., Easton, Pa.

Various other examples will be apparent to the person skilled in the artafter reading the present disclosure without departing from the spiritand scope of the invention. It is intended that all such other examplesbe included within the scope of the appended claims.

What is claimed is:
 1. A compound of the formula ##STR15## whereinW=OCH₂ Ph or OMe.
 2. A compound of the formula ##STR16## wherein W=OCH₂Ph or OMe.
 3. A compound of the formula ##STR17##